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(11) | EP 0 421 733 B1 |
| (12) | EUROPEAN PATENT SPECIFICATION |
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| (54) |
Amphotericin B lipid complex Amphotericin-B-Lipidkomplex Complexe lipidique d'amphotéricine B |
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| Note: Within nine months from the publication of the mention of the grant of the European patent, any person may give notice to the European Patent Office of opposition to the European patent granted. Notice of opposition shall be filed in a written reasoned statement. It shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention). |
[0001] Amphotericin B is a broad spectrum polyene antifungal agent that has been shown to provide effective treatment for most mycotic infections. A complete description of this antibiotic can be found in U.S. 2,908,611. However, the utility of amphotericin B has been limited by its toxicity. Long term therapy is often associated with renal toxicity and anemia, and its acute toxicity include fever, chills, nausea and vomiting. In order to overcome this toxicity, a number of approaches have been developed. These generally include initiation of therapy with graded doses of amphotericin B to determine tolerance and close monitoring of renal function. Amphotericin B is currently available as a collodial suspension for administration by infusion. Use of sodium desoxycholate is necessary to achieve a colloidal suspension of amphotericin B which is otherwise insoluble or poorly soluble in agueous solution. It is believed, when given in vivo amphotericin B is rapidly bound to lipoproteins and red blood cell membranes.
[0002] Recently, Lopez-Berestein et al. have reported a new approach to the formulation of amphotericin B which results in lower toxicity while retaining or improving on its efficacy. This approach involves the encapsulation of amphotericin B in liposomes. These results have been reported in the following articles:
1. Lopez-Berestein, G., Mehta, R., Hopfa, R.L., et al. Treatment and Prophylaxis of Disseminated Infection Due to Candida albicans in Mice with Liposome-Encapsulated Amphotericin B. Brit. J. Infect. Dis. 147: 939-944; 1983.
2. Lopez-Berestein, G., Fainstein, V., Hopfer, R.L., et al. Liposomal Amphotericin B for the Treatment of Systemic Fungal Infections in Patients with Cancer: A Preliminary Report. J. Inf. Dis. 4: 704-710; 1985.
3. Lopez-Berestein, G., Bodey, G.P., Frankel, L.S., Mehta, R. Treatment of Hepatosplenic Fungal Infections with Liposomal Amphotericin B. Br. J. Clin. Oncol. 5: 310-317: 1987.
4. Lopez-Berestein et al. Protective effect of liposomal amphotericin B against Candida albicans infections in mice: Cancer Drug Delivery 2, 183-189(1985).
[0003] Treatment of Candida infections by an amphotericin B lipid complex in mice is also disclosed in 89th Annual Meeting of the American Society for Microbiology 1989, New Orleans, LA, 14th-18th May 1989, page 7, abstract A-41.
[0004] Most recently U.S. patent 4,663,167 has been issued and describes compositions of amphotericin B encapsulated within a liposome consisting essentially of dimyristoyl phosphatidylcholine and dimyristoyl phosphatidylglycerol in an approximately 7 to 3 molar ratio.
[0005] Also, European Patent Application EP-A-282405 describes compositions of lipids and amphotericin B which have a higher ratio of amphotericin B to lipids.
[0006] Up until now, systemic fungal infections in man have required prolonged, multiple treatments with the appropriate antifungal agents. Also, with increased use of immunosuppressive agents in organ transplantation and more aggressive chemotherapy in cancer patients, infections caused by opportunistic pathogens like Candida are becoming more prevalent.
[0007] Recently, fluconazole has received approval in England for one day therapy for the treatment of Candida vaginitis.
[0008] It has now been discovered that an amphotericin B lipid complex (ABLC) can be used to treat systemic Candida infections with a single dose administration. These include in-line catheter related candidemia, non-invasive Candida esophagitis and mucosal candidiasis. The amphotericin B lipid complex has relatively high ratios of amphotericin B to lipids as described in European Patent Application EP-A-282405.
[0009] A single dose administration makes the taking of ABLC extremely simple by eliminating the need for multiple administration.
[0010] A single dose administration of ALBC to treat Candida infections will provide an effectiveness that is at least equivalent or even greater than that achieved with the use of multiple doses of amphotericin B when the toxicity of amphotericin B is considered.
[0011] Single dose administration reduces the cost of therapy which can be high due to the long duration of treatment with multiple doses.
[0012] Single dose administration reduces the probability of side-effects occurring by reducing the amount of amphotericin B necessary to treat Candida infection.
[0013] Fungal infections in man in general and Candida infections specifically require prolonged, multiple treatments with the appropriate antifungal agent. This invention provides a method of treating Candida infections in mammals which require only one dose. This is possible because of the unique properties of the amphotericin lipid complex.
[0014] The amphotericin B lipid complex of this invention is prepared by dissolving amphotericin B in DMSO, filtering and sterilizing by further filtration. This filtered and sterilized solution is aseptically combined with a filter-sterilized solution of Dimyristoyl phosphatidylcholine and, Dimyristoyl phosphatidylglycerol in methylene chloride in a sterile mixing vessel. Filter-sterilized sodium chloride injection (normal saline) is added to the above mixture in order to hydrate the complex. The methylene chloride portion of the mixture is removed by evaporation. The remaining sterile suspension of ABLC complex in saline, with residual DMSO, is further diluted with sterile normal saline for ease of handling. The suspension is aseptically milled via a sterilized colloid mill until the bulk of the product is below five microns in particle size. The milled suspension is aseptically filtered through ceramic tangential filters of five micron and 1.2 micron pore size in order to recover the fraction of drug-lipid complex between one and five microns.
[0015] The ABLC formulations described in this invention can be used to treat various fungal-related diseases in mammals such as in-line catheter related candidemia, non-invasive Candida esophagitis and mucosal candidiasis by a single-dose administration. For combating fungal infections in mammals a formulation of this invention can be administered as a single dose in an amount of about 0.5 mg/kg to about 10 mg/kg. The preferred mode of administration is the intravenous route. The following examples illustrate that a single dose administration is effective in treating systemic candidiasis in mice.
Example 1
[0016] The following ABLC composition was prepared in the manner described above.
| Amphotericin B | 5.0 mg |
| Dimyristoyl phosphatidylcholine (DMPC) | 4.6 mg |
| Dimyristoyl phosphatidylglycerol (DMPG) | 2.0 mg |
| Sodium Chloride | 9.0 mg |
| Water for Injection, enough to make | 1.0 ml |
Example 2
[0017] Female Swiss Webster mice were infected intraveneously with 5 x 10⁶ blastospores of C. albicans SC 5314 and treated one time with either ABLC or amphotericin B, five hours later. The amphotericin B was diluted in 5% dextrose for intravenous injection into the experimental mice. The ABLC formulation was also administered intravenously. The results are reproduced in Table 1 appearing below:
| Preparation | ED₅₀(mg AMB/kg) | |
| Survival | Clearance | |
| ABLC | 1.16 (1.07 - 1.28) | 1.93 (1.19 - 2.8) |
| Amphotericin B | 0.46 (0.36 - 0.54) | 0.7 (0.54 - >0.8) |
[0018] Table 1 represents a range of three experiments. As noted in Table 1, ABLC is efficacious with a survival ED₅₀ of 1.16 mg/kg of ABLC and a range of from 1.07 to 1.28 for the three experiments represented compared to 0.46 for amphotericin B, range from 0.36 to 0.54. When efficacy is based on kidney clearance, ABLC is still efficacious with an ED₅₀ of 1.93 compared to 0.7 with amphotericin B.
1. Use of Amphotericin B Lipid Complex for the manufacture of a medicament for the treatment
of systemic fungal infections in mammals in a single administration.
2. Use according to claim 1 wherein the amount to be administered is 0.5 mg/kg to about
10 mg/kg.
3. Use according to claim 1 wherein the fungal infection is inline catheter related candidemia.
4. Use according to claim 1 wherein the fungal infection is Candida esophogitis.
5. Use according to claim 1 wherein the fungal infection is mucosal candidiasis.
1. Verwendung von Amphotericin B-Lipidkomplex zur Herstellung eines Medikamentes zur
Behandlung von systemischen fungalen Infektionen bei Säugern in einer einmaligen Verabreichung.
2. Verwendung nach Anspruch 1, worin die zu verabreichende Menge 0,5 mg/kg bis etwa 10
mg/kg beträgt.
3. Verwendung nach Anspruch 1, worin die fungale Infektion Dauerkatheter-Candidämie (in-line
katheter related candedemia) ist.
4. Verwendung nach Anspruch 1, worin die fungale Infektion Candida Ösophagitis ist.
5. Verwendung nach Anspruch 1, worin die fungale Infektion mucosale Candidiasis ist.
1. Utilisation de complexe lipidique d'amphotéricine B pour la préparation d'un médicament
pour le traitement d'infections fongiques générales chez les mammifères en une administration
unique.
2. Utilisation selon la revendication 1, dans laquelle la quantité à administrer est
de 0,5 mg/kg à environ 10 mg/kg.
3. Utilisation selon la revendication 1, dans laquelle l'infection fongique est une candidémie
se rapportant à un cathéter en place.
4. Utilisation selon la revendication 1, dans laquelle l'infection fongique est une Candida
esophagitis.
5. Utilisation selon la revendication 1, dans laquelle l'infection fongique est une candidose
des muqueuses.